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This study evaluated the effects of dietary fermented tuna by‐product meal (FTBM) in juvenile olive flounder, Paralichthys olivaceus. Five diets were formulated to replace fishmeal (FM) with FTBM at 0% (FTBM0), 12.5% (FTBM12.5), 25.0% (FTBM25), 37.5% (FTBM37.5), or 50% (FTBM50). After 8 wk, weight gain, specific growth rate, and feed efficiency of fish fed FTBM0 and FTBM12.5 diets were significantly higher than fish fed the other diets (P < 0.05). Also, mean cumulative survival rates (%) of fish fed the FTBM0 and FTBM12.5 diets were significantly higher than those fed FTBM50 diet at Day 9 postchallenge with Edwardsiella tarda (P < 0.05). Protein efficiency ratio of fish fed FTBM0 and FTBM12.5 diets was significantly higher (P < 0.05) than fish fed diets FTBM37.5 and FTBM50. Broken‐line regression analysis of weight gain showed an optimal FM replacement level of 10.65% with FTBM. Therefore, the optimal dietary inclusion of FTBM in juvenile olive flounder diets could be greater than 10.65% but less than 12.5% without any adverse physiological effects on fish health.